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Differentiation of Human Wharton Jelly Mesenchymal Stem Cells into Germ-Like Cells; emphasis on evaluation of Germ-long non-coding RNAs

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Abstract

Background

The proliferation and differentiation of stem cells into Germ-Like Cells (GLCs) is mediated by several growth factors and specific genes, of which some are related to long non-coding RNAs (lncRNAs). We have developed a modified differentiation process and identified a panel of GermlncRNAs related to GLCs.

Methods

Human Wharton Jelly Mesenchymal Stem Cells were treated with 25 ng/ml Bone Morphogenetic Protein (BMP)-4 and 10− 5 M all-trans retinoic acid to differentiate them into germ-like cells. To confirm the differentiation, changes in the expression of Oct-4, C-kit, Stella, and Vasa genes were assessed using quantitative Real-Time PCR (qPCR) and immunocytochemistry. QPCR was also used before and after differentiation to evaluate the changes in a lncRNA panel, using a 96-well array. Statistical analysis of the data was performed by SPSS 21.

Results

After 21 days of induction, the HWJ-MSCs derived germ-like cells were formed. Also, qPCR and immunocytochemistry showed that the pluripotent Oct4 marker was expressed in the undifferentiated HWJ-MSCs, but its expression gradually decreased in the differentiated cells. C-kit was expressed on days 7, 14, and 21 of differentiation. Both GLC markers of Stella and Vasa genes/proteins were present only in differentiated cells. Of the 44 lncRNA genes array, 36 of them showed an increase and eight genes showed a decrease.

Conclusion

Our study showed that BMP4 and RA are effective in inducing HWJ-MSCs differentiation into GLCs. In addition, our study for the first time showed changes in the lncRNAs expression during the differentiation of HWJ-MSCs into GLCs by using BMP4 and RA.

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Acknowledgements

Thanks to Dr. Mostafa Latifpour and his colleagues to provide the Human Wharton Jelly Mesenchymal Stem Cells as a gift.

Funding

This study was derived from a thesis for an M.Sc. degree in the field of medical biotechnology (Grant or Code Number: 950214021) at Gorgan School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran.

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Authors and Affiliations

  1. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran

    Samira Ghasemi

  2. Stem Cell Research Center, Golestan University of Medical Sciences, Gorgan, Iran

    Mohammad Shafiee & Mohsen Saeedi

  3. Division of Medical Education, Brighton and Sussex Medical School, Brighton, UK

    Gordon A. Ferns

  4. Department of Immunology, BuAli Research Institute, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Jalil Tavakol-Afshari

  5. Persian Cohort Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Sara Raji

  6. Department of Laboratory Sciences, School of Paramedical Sciences, Mashhad University Of Medical Sciences, Mashhad, Iran

    Naser Mobarra

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  1. Samira Ghasemi
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Contributions

SGH: Performing experiments, acquisition of data, analysis and interpretations of data, manuscript drafting, revision of the manuscript. MSH: Contribution to study design. GAF: Contribution to data analysis and manuscript drafting. JTA: Contribution to the evaluation of experiments and manuscript drafting. MS: Contribution to study design. SR: Contribution to manuscript revision. NM: Study design and concept, participation in literature bibliography, data acquisition and analysis, manuscript drafting, and critical revision of the manuscript. All authors read and approved the final version of the manuscript.

Corresponding author

Correspondence to Naser Mobarra.

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Ethical approval

The study was approved by the Committee of Ethics, Golestan University of Medical Sciences (Code of Ethics: IR.GOUMS.REC.1394.34).

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The authors declare that they have no conflicts of interest.

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Ghasemi, S., Shafiee, M., Ferns, G.A. et al. Differentiation of Human Wharton Jelly Mesenchymal Stem Cells into Germ-Like Cells; emphasis on evaluation of Germ-long non-coding RNAs. Mol Biol Rep 49, 11901–11912 (2022). https://doi.org/10.1007/s11033-022-07961-6

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